Amphiphysin I Double Nickase Plasmid (h): sc-403904-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Amphiphysin I Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- Amphiphysin I Double Nickase Plasmid (h) and Amphiphysin I Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AMPH. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Amphiphysin I Antibody (G-4): sc-376402
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Amphiphysin I Double Nickase Plasmid (h) | sc-403904-NIC | 20 µg | $410.00 | |||
Amphiphysin I Double Nickase Plasmid (h2) | sc-403904-NIC-2 | 20 µg | $410.00 |
Human AMPH encodes amphiphysin I, a BAR domain–containing adaptor that shapes membranes and coordinates clathrin-mediated endocytosis by coupling dynamin-dependent vesicle fission with actin remodeling. Amphiphysin I participates in synaptic vesicle recycling and receptor internalization pathways through interactions with clathrin, AP-2, and SH3-binding partners, supporting efficient neurotransmission and membrane trafficking. Dysregulation of endocytic machinery and synaptic homeostasis involving AMPH is relevant to studies of neurodegeneration and other neurological disorders where vesicle recycling, protein turnover, and signaling receptor trafficking are perturbed. AMPH is therefore widely used as a mechanistic node for interrogating membrane curvature sensing, endosomal trafficking, and synapse-associated cellular stress responses.
Amphiphysin I Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AMPH locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AMPH. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt AMPH function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of AMPH-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.